ZIC1 suppresses cell proliferation and migration in gastric cancer. (A) Gastric cancer cell lines (AGS, MKN28, BGC823 and SGC7901) were stably transfected with pCDNA3.1-ZIC1 or pCDNA3.1 empty vector. The expression levels of ZIC1 mRNA and protein were performed by RT-PCR (upper diagram) and Western blot (low diagram). GAPDH and β-actin were used as internal controls. (B) The cell viability of gastric cancer cell line (BGC823) was determined by MTS cell proliferation assay. The asterisk indicates statistical significance (**p < 0. 01). (C) Cell migration was assessed by modified Boyden transwell chambers assays after incubation for 16 h. Cells that migrated to the bottom of the membrane were stained with DAPI. (D) The number of visible migratory cells (mean ± S.D) was estimated by counting five random high power fields (×400 magnification). These experiments were performed in triplicate. The representative data are shown. The asterisk indicates statistical significance (***p < 0.001).